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. 2021 Feb 8;40(5):e107037. doi: 10.15252/embj.2020107037

Figure 1. The replisome is inherently tolerant of a leading‐strand Tg lesion.

Figure 1

  • A
    Schematic of the ARS306 leading‐strand lesion template showing the origin of replication (Ori) and leading‐ (red) and lagging‐strand (blue) replication products. The SmaI restriction site is shown; cleavage is used to reduce heterogeneity in product sizes for denaturing gel analysis.
  • B
    Schematic of the uncoupled fork and uncoupled product generated by persistent stalling of leading‐strand synthesis.
  • C, D
    Standard replication assays on undamaged (UD), thymine glycol (Tg) and cyclobutane pyrimidine dimer (CPD) templates in the presence (C) and absence (D) of Pol η, Pol ζ, Rev1, Rad6–Rad18, Uba1 and ubiquitin (TLS machinery). Reactions contained 2.5 nM Pol δ.